HPLC法在国内外药典中的应用与比较课件

1、HPLC法在国内外药典中的应用与比较2022/10/10HPLC法在国内外药典中的应用与比较HPLC法在国内外药典中的应用与比较2022/10/9HPL汇报的主要内容 一、简述 二、高效液相色谱仪 三、系统适用性 四、色谱条件的调整 五、HPLC的应用及方法开发2022/10/10HPLC法在国内外药典中的应用与比较2汇报的主要内容 一、简述2022/10/9HPLC法在国内外一、简述中国药典2010年版二部共收载2271个品种，新增品种330个，修订品种1500个，涉及HPLC检测项目的品种有1291个，占总品种的57%，其中新增/修订926个。HPLC法在整个药典品种的检验中占有重要地位。

2、2022/10/10HPLC法在国内外药典中的应用与比较3一、简述中国药典2010年版二部共收载2271个品种，新High Performance Liquid Chromatography简称HPLC，开始应用于20世纪60年代后期，现已趋于成熟，广泛应用于医药、生化、天然产物主要组分分析，以及食品、化妆品分析，环境分析，农业分析，石油化工分析等。优点：高压+高速+高效+高灵敏度一、简述2022/10/10HPLC法在国内外药典中的应用与比较4High Performance Liquid ChromaCHP(2010)：中国药典2010年版USP：United Stated Pharmac

3、opoeia EP：European PharmacopoeiaJP：Japanese Pharmacopoeia一、简述2022/10/10HPLC法在国内外药典中的应用与比较5CHP(2010)：中国药典2010年版一、简述2022定义CHP(2010)附录V D：HPLC系采用高压输液泵将规定的流动相泵入装有填充剂的色谱柱，对供试品进行分离测定的色谱方法。USP(32)：High-pressure liquid chromatography (HPLC), sometimes called high-performance liquid chromatography, is a sepa

4、ration technique based on a solid stationary phase and a liquid mobile phase. Separations are achieved by partition, adsorption, or ion-exchange processes, depending upon the type of stationary phase used.2022/10/10HPLC法在国内外药典中的应用与比较6定义CHP(2010)附录V D：HPLC系采用高压输液泵将定义EP(6.0) 2.2.29：Liquid chromatograp

5、hy is a method of chromatographic separation based on the difference in the distribution of species between two non-miscible phases, in which percolates through a stationary phase contained in a column.2022/10/10HPLC法在国内外药典中的应用与比较7定义EP(6.0) 2.2.29：Liquid chroma定义JP(XV)2.01：Liquid chromatography is a

6、 method to develop a mixture injected into a column prepared with a suitable stationary phase by passing a liquid as a mobile phase through the column, in order to separate the mixture into its components by making use of the difference of retention capacity against the stationary phase, and to dete

7、rmine the components.2022/10/10HPLC法在国内外药典中的应用与比较8定义JP(XV)2.01：Liquid chromatogr二、高效液相色谱仪基本组成：2022/10/10HPLC法在国内外药典中的应用与比较9二、高效液相色谱仪基本组成：2022/10/9HPLC法在国泵CHP(2010)USP(32)EP(6.0)JP(XV) 未解释满足等度和梯度洗脱;压力：5000psi或更高;最大可达10ml/min可满足等度或梯度洗脱只强调了恒定速率泵的种类很多，目前应用最多的是柱塞往复泵（恒流泵）2022/10/10HPLC法在国内外药典中的应用与比较10泵CHP

8、(2010)USP(32)EP(6.0)JP(XV)进样器CHP(2010)USP(32)EP(6.0)JP(XV)未解释微量进样器定量环自动进样器手动 / 自动强调重复性好即可2022/10/10HPLC法在国内外药典中的应用与比较11进样器CHP(2010)USP(32)EP(6.0)JP(X色谱柱分离的核心CHP(2010)附录：1.正相：硅胶柱； 反相： C-18柱、C-8柱；2.粒径：普通310m； 2m（亚-2m仅能用于UPLC）；3.温度要求：以硅胶为载体的通常 40；不宜超过60 ；4.pH要求：28；8，硅胶溶解以5m最为常见2022/10/10HPLC法在国内外药典中的应用

9、与比较12色谱柱分离的核心CHP(2010)附录：2，化学键合相脱色谱柱技术发展很快，新填料新技术也不断涌现，给我们提供了更多的选择。Gemini-NX C18柱：色谱柱乙烷交联，抵抗高pH侵蚀多点配体键合，抵抗低pH下配体水解pH112稳定！Kinetex C18柱：通过核-壳颗粒技术，使颗粒减小到2.6m，而无需高压即可达到提高速度、分离度和灵敏度的效果。2022/10/10HPLC法在国内外药典中的应用与比较13色谱柱技术发展很快，新填料新技术也不断涌现，给我们提供了更多色谱柱Zorbax StableBond柱：采用了较大的二异丁基(SB-C18)或二异丙基(SB-C8、SB-C3、S

10、B-Phenyl、SB-CN、SB-Aq)侧链基团和空间位阻，避免了在低pH条件下的水解破坏，其pH1.08.0，温度上限也可达到8090，甚至在100%水相中也有出色的表现。2022/10/10HPLC法在国内外药典中的应用与比较14色谱柱Zorbax StableBond柱：2022/10/色谱柱Zorbax Extend C18柱：采用独特的双配位C18-C18键合技术，使的在高pH条件下使用硅胶基色谱柱成为可能，在pH211.5的范围内是稳定的。2022/10/10HPLC法在国内外药典中的应用与比较15色谱柱Zorbax Extend C18柱：2022/10/USP(32):Sta

11、tionary phases for modern, reverse-phase liquid chromatography typically consist of an organic phase chemically bound to silica or other materials.1. Particles：usually 3 to 10 m in diameter；may range up to 50 m or more for preparative columns. 2. Internal diameters：usually 2 to 5mm for analytical se

12、paration，lager for preparative chromatography.3. Temperature: only rarely above 60 ；4.根据填料不同，USP在Chromatographic reagents in the Reagents, Indicators, and Solutions section列出了USP-NF所用到的不同类型的柱子，如下所示:2022/10/10HPLC法在国内外药典中的应用与比较16USP(32):Stationary phasesC-18硅胶柱C-8NH2柱阳离子交换柱（SCX）CN柱苯基柱阴离子交换柱（SAX）2022/

13、10/10HPLC法在国内外药典中的应用与比较17C-18硅胶柱C-8NH2柱阳离子交换柱CN柱苯基柱阴离子交L57L67未列出2022/10/10HPLC法在国内外药典中的应用与比较18L57L67未列出2022/10/9HPLC法在国内外药典EP(6.0) 2.2.29： 大部分分离机制都是基于以化学键合硅胶作为固定相，极性溶剂作为流动相的色谱条件。而化学键合相的性质往往决定了色谱系统的分离性能。2022/10/10HPLC法在国内外药典中的应用与比较19EP(6.0) 2.2.29： 大部分分离机制都是基于以化学particle size：310m internal diameters：

14、prescribed in the monograph, eg:pH：silica based reversed-phase columns are considered to be stable in mobile phases having an apparent pH in the range 2.0 to 8.0.temperature: not be heated above 60 Special：EP(6.0) 2.2.29：stationary phase degradetioncomposition of the mobile phase2022/10/10HPLC法在国内外药

15、典中的应用与比较20particle size：310m EP(6.0) 2EPMannitol Assay:2022/10/10HPLC法在国内外药典中的应用与比较21EPMannitol Assay:2022/10/9HPLCJP(XV)：A column with a stationary phase chemically bound on the inside wall instead of the column packed with the packing material may be used.2022/10/10HPLC法在国内外药典中的应用与比较22JP(XV)：A col

16、umn with a station检测器CHP(2010):UVDADFLDECDELSDRIDMS优点：灵敏度高、噪音低、线性范围宽、对流速和温度的波动不灵敏，适用于梯度洗脱及制备色谱。 缺点：只能检测有紫外吸收的物质，流动相的选择有一定限制，流动相的截止波长必须小于检测波长。 适用范围：大多数有紫外吸收的化合物。优点：通用型检测器，对各种物质有几乎相同的响应。缺点：灵敏度相对较低，流动相必须是挥发性的，不能用非挥发性的缓冲盐及表面活性剂。适用范围：适用于挥发性低于流动相的组分，主要用于糖类、高级脂肪酸、磷脂、维生素、氨基酸、甘油三酯、皂苷及甾体等等无紫外吸收或紫外末端吸收的化合物的检测。

17、 优点：灵敏度高、选择性好，是微量组分和体内药物分析常用的检测器之一。 缺点：只适用于能够产生荧光的物质的检测，适用范围不如紫外检测器。影响因素较多。 适用范围：具有天然荧光的物质或通过荧光衍生化变成荧光衍生物后测定，主要用于氨基酸、多环芳烃、维生素、甾体化合物及酶等的检测。优点：灵敏度很高，尤其适用于痕量组分分析。缺点：干扰比较多，对温度和流速的变化比较敏感。适用范围：应用范围广，凡具氧化还原活性的或经衍生化后具氧化还原活性的物质。优点：灵敏度高，选择性好，能同时给出组分的结构信息。缺点：响应信号受离子化效率限制，仪器较为昂贵，流动相必须是挥发性的，不能用非挥发性的缓冲盐及表面活性剂。适用范

18、围：组分的结构鉴别，微量及痕量组分的分析，药物代谢分析等。优点：通用型检测器，只要组分的折光率与流动相的折光率有足够的差别就能检测。缺点：灵敏度低、受环境温度、流量及流动相组成等波动的影响大，一般不能用于梯度洗脱。适用范围：RID为通用型检测器，适用于无紫外吸收化合物的分析，如糖类分析。2022/10/10HPLC法在国内外药典中的应用与比较23检测器CHP(2010):优点：灵敏度高、噪音低、线性范围宽USP and EPUSP(32)EP(6.0)Fixed, variable and multi-wavelength detectors are widely available.UV、R

19、ID、FLD、ECD.New detectors continue to be developed in attempts to overcome the deficiencies of those being used.UV/Vis spectrophotometers, including diode array detectors, are the most commonly employed detectors.2022/10/10HPLC法在国内外药典中的应用与比较24USP and EPUSP(32)EP(6.0)Fixed,流动相CHP(2010)反相系统首选甲醇-水系统（采用紫

20、外末端波长检测，首选乙腈-水系统）缓冲盐：少用，尽可能低反相色谱系统，C-18柱，有机相比例应不低于5%。USP(32)High-purity reagents and “HPLC grade”organic solventswater: low condutivity and low UV absorptionEP(6.0)For nomal-phase chromatography, less polar solvents are employed. Water is to be strictly controlled!0.45mdegassed by sparing with heliu

21、m,sonication or using on-line membrane/vacuumAdjustment of the pH,is effected using only the aqueous component of the mobile phase and not the mixture.2022/10/10HPLC法在国内外药典中的应用与比较25流动相CHP(2010)反相系统首选甲醇-水系统（采用紫外末系统适用性System Suitability2022/10/10HPLC法在国内外药典中的应用与比较26系统适用性2022/10/9HPLC法在国内外药典中的应用与定义CHP(

22、2010):色谱系统的适用性试验通常包括理论板数、分离度、重复性和拖尾因子四个参数，其中，分离度和重复性尤为重要。ICH definition: System suitability testing is an integral part of many analytical procedures. The tests are based on the concept that the equipment, electronics, analytical operations and samples to be analyzed constitute an integral system th

23、at can be evaluated as such. 2022/10/10HPLC法在国内外药典中的应用与比较27定义CHP(2010):色谱系统的适用性试验通常包括理论板数DefinitionEP (6.0): The system suitability tests represent an integral part of the method and are used to ensure adequate performance of the chromatographic system. The various components of the equipment employ

24、ed must be qualified and be capable of achieving the precision required to conduct the test or assay. USP(32): System suitability tests are an integral part of gas and liquid chromatographic methods. They are used to verify that the resolution and reproducibility of the chromatographic system are ad

25、equate for the analysis to be done.2022/10/10HPLC法在国内外药典中的应用与比较28DefinitionEP (6.0): The systemDefinitionUSP(32): No sample analysis is acceptable unless the requirements of system suitablity have been met. Sample analyses obtained while the system fails requirements are unacceptable. System suitabi

26、lity must be demonstrated throughout the run by injection of an appropriate control preparation at appropriate intervals. Wherever there is a significant change in equipment or in a critical reagent, suitability testing should be performed before the injection of samples.2022/10/10HPLC法在国内外药典中的应用与比较

27、29DefinitionUSP(32): No sample 理论板数评价色谱柱的重要指标。CHP(2010) *USP(32) *EP(6.0)JP(XV)n=16(tR/W)2 n=5.54(tR/Wh/2)2N=16(t/W)2N=5.54(t/Wh/2)2N=5.54(tR/Wh)2Wh：半高峰宽N=5.54(tR/W0.5h)2W0.5h：半高峰宽 影响因素：固定相、柱温、流动相和保留时间。*有争议时，以峰宽（W）计算结果为准2022/10/10HPLC法在国内外药典中的应用与比较30理论板数评价色谱柱的重要指标。CHP(2010) *USP(分离度（R）衡量色谱系统效能的关键指标！

28、 2022/10/10HPLC法在国内外药典中的应用与比较31分离度（R）衡量色谱系统效能的关键指标！ 2022/10/9分离度（R）CHP(2010)*R=2(tR2-tR1)/(W1+W2)R=2(tR2-tR1)/1.70(W1,h/2+W2,h/2)除另有规定外，应大于1.5 USP(32) *# EP(6.0) # JP(XV)* 有争议时，以峰宽（W）计算结果为准；# described in individual monograph2022/10/10HPLC法在国内外药典中的应用与比较32分离度（R）CHP(2010)*R=2(tR2-tR1)/(USP(32)Chromato

29、graphic system The liquid chromatograph is equipped with a 275-nm detector and a 4.6-mm 15-cm column that contains packing L1 the resolution, R, between the impurity C and famotidine peaks is not less than 1.3; the resolution, R, between the famotidine and impurity D peaks is not less than 1.3;Famot

30、idine TabletsAssary 2022/10/10HPLC法在国内外药典中的应用与比较33USP(32)Chromatographic system EP(6.0)左甲状腺素钠含量测定：左羟丙哌嗪对映体纯度：2022/10/10HPLC法在国内外药典中的应用与比较34EP(6.0)左甲状腺素钠含量测定：左羟丙哌嗪对映体纯度：2重复性 评价连续进样中，色谱系统响应值的重复性能。 CHP(2010):外标法：对照品溶液，连续进样5次，峰面积RSD不得过2.0%。内标法：配制相当于80%、100%、120%的对照品溶液，加入内标溶液，分别至少进样2次，计算平均校正因子的RSD不得过2.0%

31、 。2022/10/10HPLC法在国内外药典中的应用与比较35重复性 评价连续进样中，色谱系统响应值的重复性能。202重复性 USP(32):Unless otherwise specified in the individual monograph , data from five replicate injections of the analyte are used to calculate the relative standard deviation, SR, if the requirment is 2.0% or less; data from six replicate in

32、jections are used if the relative standard deviation requirment is more than 2.0%. 2022/10/10HPLC法在国内外药典中的应用与比较36重复性 USP(32):2022/10/9HPLC法在国内重复性 EP(6.0):The repeatability of response is expressed as an es timated percentage relative standard deviation(Sr(%) of a consecutive series of measurements f

33、or NOT fewer than 3 injections or applications of a reference solution.mean of individual valuesindividual values expressed as peak area, peak height, or ratio of areas by the interal standardisation methodnumber of individual values2022/10/10HPLC法在国内外药典中的应用与比较37重复性 EP(6.0):mean of individu重复性 EP(6.

34、0):Unless otherwise prescribed, the maximum permitted RSD does not exceed the appropriate value given in table.Number of individual injections3456B(percent)Maximum permitted relative standard deviation2.00.410.590.730.852.50.520.740.921.063.00.620.891.101.272022/10/10HPLC法在国内外药典中的应用与比较38重复性 EP(6.0):

35、Number of indivi重复性NOTE: This requirement does not apply to tests for related substances.upper limit given in the definition of the individual monograph minus 100%contant(0.349)number of replicate injections of the reference solution (3n6)90% probability level，n-1 degrees of freedomEP(6.0): In an as

36、say of an active substance where the value is 100 percent for a pure substance, the maximum permitted (Sr(%)max) for defined limits is calculated using the following equation:2022/10/10HPLC法在国内外药典中的应用与比较39重复性NOTE: This requirement does重复性 JP(XV): 公式与EP相同。重复次数及限度在各论中要求。例： Ritodrine Hydrochloride Rela

37、ted subsances System suitability Test for required detectability: . System performance:. System repeatability: When the test is repeated 6 times with .，the relative standard deviation of the peak areas of deferoxamine is not more than 3.0%.2022/10/10HPLC法在国内外药典中的应用与比较40重复性 JP(XV): 公式与EP相同。2022/10/拖尾

38、因子（对称因子）用于评价色谱峰的对称性2022/10/10HPLC法在国内外药典中的应用与比较41拖尾因子（对称因子）用于评价色谱峰的对称性2022/10/9拖尾因子T（or Symmetry factor）CHP(2010)T=W0.05h/2d1应符合个论项下的规定除另有规定外，峰高法定量时T应在0.951.05之间USP(32)T=W0.05/2fEP(6.0)As=W0.05h/2dAn As value of 1.0 signifies symmetry. When As 1.0, the peak is tailing. When As 1.0, the peak is front

39、ing.In a related substances test or assay, for a peak in the chromatogram obtained with a reference solution used for quantification, the symmetry factor is 0.8 to 1.5, unless otherwise prescribed.JP(XV)S=W0.05h/2f2022/10/10HPLC法在国内外药典中的应用与比较42拖尾因子T（or Symmetry factor）CHP(2USPIt is also a common pra

40、ctice to measure the Asymmetry factor as the ratio of the distance between the vertical line connecting the peak apex with the interpolated baseline and the peak front, and the distance between that line and the peak back measure at 10% of the peak height, it would be (W0.10-f0.10)/f0.10However, for

41、 the purpose of USP, only the formula presented in the Glossary of Symbols is valid. 2022/10/10HPLC法在国内外药典中的应用与比较43USPIt is also a common practicEP中系统适用性：p/v and S/NThe peak-to-valley ratio (p/v) may be employed as a system suitability criterion in a test for related substances when baseline separat

42、ion between 2 peaks is not achieved.p/v=Hp/HvFor example: Econazole related substances2022/10/10HPLC法在国内外药典中的应用与比较44EP中系统适用性：p/v and S/NThe peak-tEPThe short-term noise influences the precision of quantification.The signal-to-noise ratio is calculated using the following equation:S/N=2H/h For exampl

43、e: ketotifen hydrogen fumarate related substances2022/10/10HPLC法在国内外药典中的应用与比较45EPThe short-term noise influen色谱条件的调整Adjustment of Chromatographic Conditions2022/10/10HPLC法在国内外药典中的应用与比较46色谱条件的调整2022/10/9HPLC法在国内外药典中的应CHP(2010)明确规定：不可变的有：固定相的种类、流动相的组分、检测器类型可变的有：色谱柱内径、长度、载体粒度、流动相流速、混合流动相各组分的比例、柱温、进样量、检

44、测器的灵敏度。2022/10/10HPLC法在国内外药典中的应用与比较47CHP(2010)明确规定：2022/10/9HPLC法在国CHP(2010)与CHP(2005)不同：中国药典2010年版规定了流动相调整的限度。调整流动相组分比例时，以组分比例较低者（50）相对于自身改变量不超过30且相对于总量的改变量不超过10为限，如30相对改变量的数值超过总量的10时，则改变量以总量的10为限。 2022/10/10HPLC法在国内外药典中的应用与比较48CHP(2010)与CHP(2005)不同：中国药典20CHP(2010)例：奥美拉唑肠溶片，释放度检查，色谱条件：问题：峰型差！流动相可调范

45、围：82.5:17.5 67.5:32.5解决：70：302022/10/10HPLC法在国内外药典中的应用与比较49CHP(2010)例：奥美拉唑肠溶片，释放度检查，色谱条件：USP(32)pH of mobile phaseconcentration of saltsin bufferratio of components in mobile phaseWavelength of UV detectorinjection volumepH of the aqueous buffer within 0.2 units of the value or range specified withi

46、n 10%provided the permitted pH variation is metapply to minor components can be adjusted by 30 relativecan not exceed 10% absolute, see example:binary or ternary mixturesnot permittedcan be reduced as far as is consistent with accepted precision and detection limitscolumn lengthcolumn inner diameter

47、particle sizeflow ratecolumn temperature70%25%can be reduced by as much as 50%50%10 If adjustments of operating conditions to meet system suitability requirments are necessary, each of the following is the maxium variation that can be considered, unless otherwise directed in the monograph.2022/10/10

48、HPLC法在国内外药典中的应用与比较50USP(32)pH of mobile phaseconceUSP(32)Binary Mixtures SPECIFIED RATIO OF 50:50 Thirty percent of 50 is 15% absolute, but this exceeds the maximum permitted change of 10% absolute in either component. Therefore, the mobile phase ratio may be adjusted only within the range of 40:60

49、to 60:40. SPECIFIED RATIO OF 2:98 Thirty percent of 2 is 0.6% absolute. Therefore the maximum allowed adjustment is within the range of 1.4:98.6 to 2.6:97.4. 2022/10/10HPLC法在国内外药典中的应用与比较51USP(32)Binary Mixtures 2022/USP(32)Ternary Mixtures SPECIFIED RATIO OF 60:35:5 For the second component, 30% of

50、35 is 10.5% absolute, which exceeds the maximum permitted change of 10% absolute in any component. Therefore the second component may be adjusted only within the range of 25% to 45% absolute. For the third component, 30% of 5 is 1.5% absolute. In all cases, a sufficient quantity of the first compone

51、nt is used to give a total of 100%.Therefore, mixture ranges of 50:45:5 to 70:25:5 or 58.5:35:6.5 to 61.5:35:3.5 would meet the requirement. 2022/10/10HPLC法在国内外药典中的应用与比较52USP(32)Ternary Mixtures 2022EP(6.0)The extent to which the various parameters of a chromatographic test may be adjusted to satisf

52、y the system suitability criteria without fundamentally modifying the methods are listed below.Isocratic elution :pH of the aqueous component of the mobile phaseconcentration of salts in buffercomposition of the mobile phaseWavelengthcolumn temperature0.2pH 1.0pH (non-ionisable) within 10%Minor comp

53、onent 30% relative or 2% absolute whichever is larger.No component can be altered by more than 10% absolute （see example）no adjustment permitted10column lengthinteral diameterparticle sizeflow rateinjection volume70%25%maximum reduction of 50%no increase permitted50% a larger adjustment is acceptabl

54、e（see example）may be decreasedno increase permitted2022/10/10HPLC法在国内外药典中的应用与比较53EP(6.0)The extent to which theEP(6.0)Composition of the mobile phase: the amount of the minor solvent component may be adjusted by 30 % relative or 2 % absolute, whichever is the larger; for a minor component at 10 % of

55、 the mobile phase, a 30 % relative adjustment allows a range of 7-13% wherea 2% absolute adjustment allows a range of 8-12%, the relative value being therefore the larger;For a minor component at 5% of the mobile phase, a 30% relative adjustment allows a range of 3.5-6.5% whereas a 2% absolute adjus

56、tment allows a range of 3-7%, the absolute value being in this case the larger.No other component is altered by more than 10% absolute.2022/10/10HPLC法在国内外药典中的应用与比较54EP(6.0)Composition of the mobiEP(6.0)When column dimensions are changed,the flow rate may be ajusted as necessary using equation:F1：规定的

57、流速F2：调整后的流速l1：规定的柱长l2：实际所用的柱长d1：规定柱子的内径d2：实际所用柱子的内径2022/10/10HPLC法在国内外药典中的应用与比较55EP(6.0)When column dimensions EP(6.0)pH of the aqueous component of the mobile phaseconcentration of salts in buffercomposition of the mobile phaseWavelength of UV detectorcolumn temperatureno adjustmentpermitted no adj

58、ustment permittedthe system suitability requirements are fulfilleldthe principal peak elute within 15% of the indicated retention timeelution power is not weaker than beforeno adjustment permitted5column lengthinteral diameterparticle sizeflow rateinjection volume70%25%no adjustment permittedsame as

59、isocratic elution may be decreasedno increase permittedGradient elution：more critical than with isocratic elution2022/10/10HPLC法在国内外药典中的应用与比较56EP(6.0)pH of the aqueous conceHPLC的应用及方法开发2022/10/10HPLC法在国内外药典中的应用与比较57HPLC的应用及方法开发2022/10/9HPLC法在国内外HPLC应用类型定 义I原料药或药物制剂活性成分中主要成分的分析程序-如含量测定II原料药中杂质检查或药物制剂

60、中降解产物的分析程序，这些分析程序包括定量试验或限度试验III用于性能参数测试的分析程序-如溶出度、释放度IV鉴别试验HPLC 可以用来干什么？2022/10/10HPLC法在国内外药典中的应用与比较58HPLC应用类型定 义I原料药或药物制剂活性成分中主要成分方法开发的指导原则ICHUSP 中国药典2022/10/10HPLC法在国内外药典中的应用与比较59方法开发的指导原则ICH2022/10/9HPLC法在国内外ICHICH(人用药品注册技术要求国际协调会)三方协调指导原则 Q2A：分析方法论证的文本ICH三方协调指导原则 Q2B：方法学ICH三方协调指导原则2022/10/10HPLC